Limits...
Trimethoxy-benzaldehyde levofloxacin hydrazone inducing the growth arrest and apoptosis of human hepatocarcinoma cells.

Sun JP, Shi ZY, Liu SM, Kang YH, Hu GQ, Huangfu CS, Deng JB, Liu B - Cancer Cell Int. (2013)

Bottom Line: There was a significant increase of cytochrome c in the cytosol after 24 h of treatment with QNT11 and a decrease in the mitochondrial compartment.In addition, QNT11 increased the protein expression of Bax, caspase-9, caspase-8, caspase-3, as well as the cleaved activated forms of caspase-9, caspase-8 and caspase-3 significantly, whereas the expression of Bcl-2 decreased.The growth inhibition was in large part mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: College of Nursing, Institute of Neurobiology, Henan University, Kaifeng, China. kangyh0811@gmail.com.

ABSTRACT

Background: In order to search for new structural modification strategies on fluoroquinolones, we have designed and synthesized a series of fluoroquinolone derivatives by linking various hydrazine compounds to the C-3 carboxyl group of levofloxacin and assessed their anticancer activities. Several novel levofloxacin derivatives displayed potent cytotoxicity against the tested cancer cell lines in vitro. In the present study, we investigated the effect of 1-Cyclopropyl-6-fluoro-4-oxo-7- piperazin-1, 4-dihydro- quinoline- 3-carboxylic acid benzo [1,3] dioxol-5- ylmethylene- hydrazide (QNT11) on the apoptosis of human hepatocarcinoma cells in vitro.

Methods: The inhibition effects of QNT11 on cell proliferation were examined by MTT assay. Cell apoptosis was determined by TUNEL and DNA agarose gel electrophoresis method. The topoisomerase ΙΙ activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate. Cell cycle progression was analyzed using flow cytometry in conjunction with ethanol fixation and propidium iodide staining. Mitochondrial membrane potential (△ψm) was measured by high content screening image system. The caspase-9, caspase-8, caspase-3, Bcl-2, Bax, CDK1, Cyclin B1and cytochrome c protein expressions were detected by Western blot analysis.

Results: QNT11 showed selective cytotoxicity against Hep3B, SMMC-7721, MCF-7 and HCT-8 cell lines with IC50 values of 2.21 μM, 2.38 μM, 3.17 μM and 2.79 μM, respectively. In contrast, QNT11 had weak cytotoxicity against mouse bone marrow mesenchymal stem cells (BMSCs) with IC50 value of 7.46 μM. Treatment of Hep3B cells with different concentrations of QNT11 increased the percentage of the apoptosis cells significantly, and agarose gel electrophoresis revealed the ladder DNA bands typical of apoptotic cells, with a decrease in the mitochondrial membrane potential. Compared to the control group, QNT11 could influence the DNA topoisomerase IIactivity and inhibit the religation of DNA strands, thus keeping the DNA in fragments. There was a significant increase of cytochrome c in the cytosol after 24 h of treatment with QNT11 and a decrease in the mitochondrial compartment. Observed changes in cell cycle distribution by QNT11 treated might be caused by insufficient preparation for G2/M transition. In addition, QNT11 increased the protein expression of Bax, caspase-9, caspase-8, caspase-3, as well as the cleaved activated forms of caspase-9, caspase-8 and caspase-3 significantly, whereas the expression of Bcl-2 decreased.

Conclusions: Our results showed that QNT11 as a fluoroquinolone derivative exerted potent and selectively anticancer activity through the mechanism of eukaryotic topoisomerase II poisoning. The growth inhibition was in large part mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.

No MeSH data available.


Related in: MedlinePlus

Inhibition of cell viability by QNT11 and levofloxacin treatment in human cancer cells and BMSCs. Hep3B cells (A), SMMC-7721 cells (B), MCF-7 cells (C), HCT-8 cells (D) and BMSCs (E) were treated with various concentrations of QNT11 for 12–72 h. Hep3B cells were treated with various concentrations of Levofloxacin (F) for 12–72 h. Control cells were treated with the same volume of DMSO as a vehicle control (the final concentration of DMSO was below 0.1%). After treatment, cell viability was measured by MTT assay as described in Methods, and then calculated as a percentage of viability of the control cells. Data represent means ± SD of three independent measurements.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3716872&req=5

Figure 2: Inhibition of cell viability by QNT11 and levofloxacin treatment in human cancer cells and BMSCs. Hep3B cells (A), SMMC-7721 cells (B), MCF-7 cells (C), HCT-8 cells (D) and BMSCs (E) were treated with various concentrations of QNT11 for 12–72 h. Hep3B cells were treated with various concentrations of Levofloxacin (F) for 12–72 h. Control cells were treated with the same volume of DMSO as a vehicle control (the final concentration of DMSO was below 0.1%). After treatment, cell viability was measured by MTT assay as described in Methods, and then calculated as a percentage of viability of the control cells. Data represent means ± SD of three independent measurements.

Mentions: The cytotoxicity of QNT11 against cells was assessed using MTT cell viability assay. The cells were treated with various concentrations of QNT11 for 12,24, 48 and 72 h, resulting in a significant decrease in cell viability in a dose- and time-dependent manner (Figure 2). Within the four cancer cell lines used in this experiment, QNT11 was more effective against Hep3B cells. As shown in Figure 2A, the IC50 value for 12,24, 48 and 72 h treatment was 1.92 ± 0.19 μM (r2 = 0.9601), 2.21 ± 0.20 μM (r2 = 0.9679), 2.55 ± 0.25 μM (r2 = 0.9561) and 2.70 ± 0.22 μM (r2 = 0.9802) respectively. For SMMC-7721 cells, the IC50 value for 12, 24, 48 and 72 h treatment obtained was 2.16 ± 0.28 μM (r2 = 0.9109), 2.38 ± 0.22 μM (r2 = 0.9408), 2.68 ± 0.25 μM (r2 = 0.9427) and 3.06 ± 0.19 μM (r2 = 0.9264), respectively. For MCF-7 cells, the IC50 was 2.93 ± 0.23 μM (r2 = 0.9640), 3.17 ± 0.26 μM (r2 = 0.9212), 3.11 ± 0.32 μM (r2 = 0.9490) and 3.68 ± 0.25 μM (r2 = 0.9624) for 12, 24, 48 and 72 h treatment respectively. For HCT-8 cells, the IC50 value for12, 24, 48 and 72 h treatment obtained was 2.12 ± 0.22 μM (r2 = 0.9260), 2.79 ± 0.22 μM (r2 = 0.9547), 2.86 ± 0.25 μM (r2 = 0.9139) and 3.46 ± 0.32 μM (r2 = 0.9174), respectively. In contrast, QNT11 had weak cytotoxicity against BMSCs. The IC50 value for 12, 24, 48 and 72 h treatment was 8.13 ± 0.55 μM (r2 = 0.9574), 7.46 ± 0.50 μM (r2 = 0.9034), 7.90 ± 0.55 μM (r2 = 0.9414) and 12.05 ± 0.69 μM (r2 = 0.8993), respectively.


Trimethoxy-benzaldehyde levofloxacin hydrazone inducing the growth arrest and apoptosis of human hepatocarcinoma cells.

Sun JP, Shi ZY, Liu SM, Kang YH, Hu GQ, Huangfu CS, Deng JB, Liu B - Cancer Cell Int. (2013)

Inhibition of cell viability by QNT11 and levofloxacin treatment in human cancer cells and BMSCs. Hep3B cells (A), SMMC-7721 cells (B), MCF-7 cells (C), HCT-8 cells (D) and BMSCs (E) were treated with various concentrations of QNT11 for 12–72 h. Hep3B cells were treated with various concentrations of Levofloxacin (F) for 12–72 h. Control cells were treated with the same volume of DMSO as a vehicle control (the final concentration of DMSO was below 0.1%). After treatment, cell viability was measured by MTT assay as described in Methods, and then calculated as a percentage of viability of the control cells. Data represent means ± SD of three independent measurements.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3716872&req=5

Figure 2: Inhibition of cell viability by QNT11 and levofloxacin treatment in human cancer cells and BMSCs. Hep3B cells (A), SMMC-7721 cells (B), MCF-7 cells (C), HCT-8 cells (D) and BMSCs (E) were treated with various concentrations of QNT11 for 12–72 h. Hep3B cells were treated with various concentrations of Levofloxacin (F) for 12–72 h. Control cells were treated with the same volume of DMSO as a vehicle control (the final concentration of DMSO was below 0.1%). After treatment, cell viability was measured by MTT assay as described in Methods, and then calculated as a percentage of viability of the control cells. Data represent means ± SD of three independent measurements.
Mentions: The cytotoxicity of QNT11 against cells was assessed using MTT cell viability assay. The cells were treated with various concentrations of QNT11 for 12,24, 48 and 72 h, resulting in a significant decrease in cell viability in a dose- and time-dependent manner (Figure 2). Within the four cancer cell lines used in this experiment, QNT11 was more effective against Hep3B cells. As shown in Figure 2A, the IC50 value for 12,24, 48 and 72 h treatment was 1.92 ± 0.19 μM (r2 = 0.9601), 2.21 ± 0.20 μM (r2 = 0.9679), 2.55 ± 0.25 μM (r2 = 0.9561) and 2.70 ± 0.22 μM (r2 = 0.9802) respectively. For SMMC-7721 cells, the IC50 value for 12, 24, 48 and 72 h treatment obtained was 2.16 ± 0.28 μM (r2 = 0.9109), 2.38 ± 0.22 μM (r2 = 0.9408), 2.68 ± 0.25 μM (r2 = 0.9427) and 3.06 ± 0.19 μM (r2 = 0.9264), respectively. For MCF-7 cells, the IC50 was 2.93 ± 0.23 μM (r2 = 0.9640), 3.17 ± 0.26 μM (r2 = 0.9212), 3.11 ± 0.32 μM (r2 = 0.9490) and 3.68 ± 0.25 μM (r2 = 0.9624) for 12, 24, 48 and 72 h treatment respectively. For HCT-8 cells, the IC50 value for12, 24, 48 and 72 h treatment obtained was 2.12 ± 0.22 μM (r2 = 0.9260), 2.79 ± 0.22 μM (r2 = 0.9547), 2.86 ± 0.25 μM (r2 = 0.9139) and 3.46 ± 0.32 μM (r2 = 0.9174), respectively. In contrast, QNT11 had weak cytotoxicity against BMSCs. The IC50 value for 12, 24, 48 and 72 h treatment was 8.13 ± 0.55 μM (r2 = 0.9574), 7.46 ± 0.50 μM (r2 = 0.9034), 7.90 ± 0.55 μM (r2 = 0.9414) and 12.05 ± 0.69 μM (r2 = 0.8993), respectively.

Bottom Line: There was a significant increase of cytochrome c in the cytosol after 24 h of treatment with QNT11 and a decrease in the mitochondrial compartment.In addition, QNT11 increased the protein expression of Bax, caspase-9, caspase-8, caspase-3, as well as the cleaved activated forms of caspase-9, caspase-8 and caspase-3 significantly, whereas the expression of Bcl-2 decreased.The growth inhibition was in large part mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: College of Nursing, Institute of Neurobiology, Henan University, Kaifeng, China. kangyh0811@gmail.com.

ABSTRACT

Background: In order to search for new structural modification strategies on fluoroquinolones, we have designed and synthesized a series of fluoroquinolone derivatives by linking various hydrazine compounds to the C-3 carboxyl group of levofloxacin and assessed their anticancer activities. Several novel levofloxacin derivatives displayed potent cytotoxicity against the tested cancer cell lines in vitro. In the present study, we investigated the effect of 1-Cyclopropyl-6-fluoro-4-oxo-7- piperazin-1, 4-dihydro- quinoline- 3-carboxylic acid benzo [1,3] dioxol-5- ylmethylene- hydrazide (QNT11) on the apoptosis of human hepatocarcinoma cells in vitro.

Methods: The inhibition effects of QNT11 on cell proliferation were examined by MTT assay. Cell apoptosis was determined by TUNEL and DNA agarose gel electrophoresis method. The topoisomerase ΙΙ activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate. Cell cycle progression was analyzed using flow cytometry in conjunction with ethanol fixation and propidium iodide staining. Mitochondrial membrane potential (△ψm) was measured by high content screening image system. The caspase-9, caspase-8, caspase-3, Bcl-2, Bax, CDK1, Cyclin B1and cytochrome c protein expressions were detected by Western blot analysis.

Results: QNT11 showed selective cytotoxicity against Hep3B, SMMC-7721, MCF-7 and HCT-8 cell lines with IC50 values of 2.21 μM, 2.38 μM, 3.17 μM and 2.79 μM, respectively. In contrast, QNT11 had weak cytotoxicity against mouse bone marrow mesenchymal stem cells (BMSCs) with IC50 value of 7.46 μM. Treatment of Hep3B cells with different concentrations of QNT11 increased the percentage of the apoptosis cells significantly, and agarose gel electrophoresis revealed the ladder DNA bands typical of apoptotic cells, with a decrease in the mitochondrial membrane potential. Compared to the control group, QNT11 could influence the DNA topoisomerase IIactivity and inhibit the religation of DNA strands, thus keeping the DNA in fragments. There was a significant increase of cytochrome c in the cytosol after 24 h of treatment with QNT11 and a decrease in the mitochondrial compartment. Observed changes in cell cycle distribution by QNT11 treated might be caused by insufficient preparation for G2/M transition. In addition, QNT11 increased the protein expression of Bax, caspase-9, caspase-8, caspase-3, as well as the cleaved activated forms of caspase-9, caspase-8 and caspase-3 significantly, whereas the expression of Bcl-2 decreased.

Conclusions: Our results showed that QNT11 as a fluoroquinolone derivative exerted potent and selectively anticancer activity through the mechanism of eukaryotic topoisomerase II poisoning. The growth inhibition was in large part mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.

No MeSH data available.


Related in: MedlinePlus